Helmintos e seus produtos são potentes moduladores da resposta imune. Componentes de alta massa molecular do extrato de Ascaris suum (PI) suprimem a resposta anti-ovalbumina. Nas células apresentadoras de antígeno (APCs) como as dendríticas (DCs) os componentes do PI inibem a expressão de moléculas MHC-II e coestimuladoras e, assim a ativação de linfócitos TCD4+. Receptores de membrana de APCs como os do tipo Toll (TLRs) e lectina tipo-C (CLRs) reconhecem padrões moleculares de patógenos e modulam a resposta imune efetora. Assim sendo, foi estudada o papel de TLRs e CLRs expressos nas APCs na supressão induzida por PI. Pôde-se observar que o PI inibe a expressão de TLR1, 2 e 4 na fase de indução da resposta adaptativa. O efeito supressor do PI na resposta anti-OVA e na maturação de DCs é independente de TLR2 e 4, entretanto os CLRs parecem estar envolvidos nesse processo. Portanto, estes dados podem contribuir no esclarecimento dos mecanismos de ação de substâncias imunossupressoras. / Helminths and antigens derived from them are potent immunemodulators. High molecular weight components of Ascaris suum extract (PI) suppress the anti-ovalbumin response. The PI components inhibit in antigen-presenting cells (APCs), as dendritic cells (DCs), the expression of MHC-II and coestimulatory molecules and, thus the CD4 + T cells activation. APCs via membrane receptors, as the Toll like (TLRs) and C-type lectin (CLRs), recognize distinct pathogens and then promote the effector immune response. Therefore, it was studied the role of TLRs and CLRs expressed on APCs in immunesuppression induced by PI. It was observed that PI inhibited the expression of TLR1, 2, 4 and 9 in the induction phase of adaptive response. The suppressive effect of PI in anti-OVA response and DCs maturation was independent of TLR2 and TLR4, however the CLRs seem to be involved in this process. These data can contribute to clarify the action mechanisms of immunosuppressive molecules.

A presença de substâncias húmicas em águas destinadas ao abastecimento tem ocasionado diversos problemas, entre eles a formação de subprodutos halogenados, principalmente quando se emprega a pré-oxidação com cloro. Com o reagente de Fenton não há a possibilidade de formação desses indesejáveis compostos halogenados, pois o forte poder oxidativo do reagente de Fenton é devido ao radical hidroxila (OH). Aliado a isto, com o mesmo produto químico se pode ter a pré-oxidação e coagulação de compostos orgânicos, pois o reagente de Fenton consiste na aplicação de íons de Fe+2 e peróxido de hidrogênio em meio ácido, sendo o Fe+2 oxidado a Fe+3. Como esses íons formam espécies hidrolisadas que atuam como coagulantes, o reagente de Fenton pode ter a dupla função de oxidação e coagulação nos processos de tratamento. Por isso, neste trabalho foi proposta uma metodologia para emprego do reagente de Fenton como agente coagulante no tratamento de águas contendo cor elevada causada pela introdução de substâncias húmicas extraídas de turfa. Além disto, foi estudado o efeito da massa molecular das substâncias húmicas na eficiência da coagulação, floculação e flotação de águas; para isto foram preparadas 4 águas distintas apresentado a mesma cor verdadeira, mas com substâncias húmicas de diferentes massas moleculares, obtidas por fracionamento por ultrafiltração. Por meio da otimização da dosagem de coagulante e respectivo pH de coagulação e posterior construção dos diagramas de coagulação para cada água de estudo, verificou-se que a água preparada com as substâncias húmicas de menor massa molecular apresentou maior grau de dificuldade para tratamento, requerendo dosagens de coagulante (reagente de Fenton) bem mais elevadas em relação às águas preparadas com substâncias húmicas de maior massa molecular. Adicionalmente, foram realizados ensaios de filtração em areia após a flotação, procurando-se simular tratamento em ciclo completo, tendo sido constatado que com o reagente de Fenton empregado como agente coagulante, a água filtrada atendeu ao padrão de potabilidade vigente no Brasil em relação aos parâmetros medidos (cor aparente < ou = 3 uH, turbidez < 0,5 uT e ferro total residual < 0,005 mg/L). Os valores de absorvância e carbono orgânico total da água filtrada também foram muito pequenos, indicando que a formação dos subprodutos da desinfecção com cloro seriam insignificantes. / Humic substances\' presence in water destined for supply has brought many problems, such as halogenated byproducts formation, mainly when chlorine is used as preoxidant. With Fenton\'s reagent there is no formation possibility of these undesirable halogenated compounds, since the strong oxidative power of Fenton\'s reagent is due to hydroxyl radical (OH). In addition, with the same chemical product, it is possible to have the preoxidation and coagulation of organic compounds, because Fenton\'s reagent consists of Fe+2 ions and hydrogen peroxide application under acid conditions, with Fe+2 oxidized to Fe+3. Since these ions form hydrolyzed species that act as coagulants, Fenton\'s reagent can have the double function of oxidation and coagulation on treatment processes. Because of this, the current work proposed a methodology to use Fenton\'s reagent as coagulant agent in the treatment of waters having high true color caused by the introduction of humic substances extracted by peat. Besides this, humic substances molecular weight effect on coagulation, flocculation and flotation of waters was studied; for this, 4 distinct waters having the same true color were prepared, but with different humic substances molecular weights, obtained by ultrafiltration fractioning. Through optimization of coagulant dosage and respective coagulation pH and posterior construction of coagulation diagrams for each studied water, it was verified that the water prepared withthe smallest molecular weight humic substances was more difficult to treat, requiring higher coagulant (Fenton\'s reagent) dosages compared with the waters prepared with larger molecular weight humic substances. Furthermore, experiments of filtration after flotation were carried out, trying to simulate a complete cycle treatment, which verified that with Fenton\'s reagent, employed as coagulant agent, the filtered water reached the brazilian potable standards concerning the measured parameters (apparent color < ou = 3 HU, turbidity < 0.5 TU and residual total iron < 0.005 mg/L). The absorbance and total organic carbon values of filtered water were also very low, indicating that the disinfection byproducts formation with chlorine would be insignificant.

Orientador: Telma Teixeira Franco / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-02T19:11:38Z (GMT). No. of bitstreams: 1 Todorovic_Zoran_M.pdf: 2109964 bytes, checksum: a9300d59d70b5fdcbb2831ea8b7e1899 (MD5) Previous issue date: 2002 / Resumo: A biocompatibilidade, biodegradabilidade e o fato de ser um polimero catiônico bioadesivo fazem a quitosana um material atrativo para inúmeras aplicações na área biotecnológica, agrícola, farmacêutica e médica. As quito sanas com massas molares baixas e os oligômeros têm maiores perspectivas nas últimas duas áreas. O objetivo deste projeto de pesquisa foi a obtenção de polimeros com massa molar média reduzida devido à ação da enzima papaína. Empregando papaína livre e em forma imobilizada a redução da massa molar da quito sana foi estudada. Quitina foi utilizada como suporte para imobilização da papaína e para imobilização da proteína modelo albumina de soro bovino. Experimentos com todos estes materiais foram realizados para estudar os efeitos da modificação da quito sana. Os experimentos foram conduzidos em reatores batelada com pH e temperatura controlados. Soluções da quito sana (1 %) foram preparadas utilizando ácido lático ou ácido acético para solubilização. Duas técnicas para determinação da massa molar da quito sana foram utilizadas. Em primeiro lugar, a visco simetria foi empregada sendo esta a técnica mais comum para investigação de redução da massa molar. Redução da viscosidade de soluções de quitosana foi associada à diminuição da massa molar média. Outro método utilizado para a determinação de massas molares foi a cromatografia de permeação em gel, que é considerado um método relativo de determinação de massa molar, pois necessita de padrões dos polimeros para a construção de uma curva padrão. A massa molar determinada por estes dois métodos é dependente do tamanho da molécula (diâmetro). O método absoluto de determinação da massa molar com detector de espalhamento de luz também foi utilizado. Este detector conectado on-line com cromatografia de permeação em gel fornece dados absolutos da massa molar média. Luz espalhada de diferentes partes da molécula do polimero em diferentes ângulos geram o valor correto da massa molar sem utilização dos padrões. Os resultados mostraram o efeito superior na redução da massa molar utilizando papaína em comparação com outros materiais utilizados. Esta redução da massa molar média é rápida e acentuada nos primeiros minutos da reação, em comparação com a continuação do processo / Abstract: Biocompatibility, biodegradability and the fact of being bioadhesive polycationic polymer make chitosan an attractive material for various applications in agricultural and biotechnological, pharmaceutical and medical fields. Chitosans with lower molecular weights and oligomers OOve major perspectives in the last two fields. An objective of this project of research was the production of polymers with reduced molecular weight due to the action of enzyme papain. Applying papain in the free and immobilized form the reduction of molecular weight of chitosan was studied. Chitin was used as a support for the immobilization of papain and for the immobilization of model protein bovine serum albumin. Experiments with all these materiais were realized to study the effects of modification of chitosan. Experiments were run using batch reactors at controled pH and temperature. Chitosan solutions (1 %) were prepared using lactic acid or acetic acid for solubilization. Two different techniques were used for determination of molecular weight of chitosan. First viscosimetry was applied as the most common technique used for investigation of reduction of molecular weights. Reduction of viscosity of chitosan solutions is associated with the reduction of molecular weight. The other method used for determination of molecular weights is gel permeation chromatography toot is considered a relative method of determination of molecular weights as it requires polymeric standards for the construction of the standard curve. Molecular weight determined by these two methods is dependant of the molecular size (diameter). Absolute method of determination of molecular weights through measurements with detector of light scattering was aIso applied in this studies. This detector connected on-line with gel penneation chromatography provides us with the absolute molecular weights. This method does not require standards measuring the light scattered from different parts of the polymeric molecule in different angles generating in this way the correct value of molecular weight. Results show the superior effect of papain on the reduction of molecular weight compared to the other materiais used. This reduction of molecular weight is fast, pronounced in the :fust minutes of the reaction in comparison with the continuation of the process / Mestrado / Desenvolvimento de Processos Químicos / Mestre em Engenharia Química

Les propriétés hydrophobes des polymères extracellulaires (PEC) exercent l’influence profonde sur les propriétés de la surface cellulaire. Cependant, de nombreux facteurs tels que les méthodes d'extractions, le type de substrat influencent les caractéristiques des PEC et les informations concernant des caractéristiques hydrophobes sont rarement documentées. L'objectif principal de cette étude est de développer une méthode appropriée pour étudier l'hydrophobicité des PEC, puis d'étudier les caractéristiques hydrophobes des PEC.Le fractionnement hydrphobe par la résine Supelite™ DAX-8 a d'abord été appliqué sur les PEC extraits de boues granulaires anaérobies, deux conditions de pH d'élution (pH 2 et 5) ont été testées. L'impact de sept méthodes d'extraction sur les caractéristiques hydrophobes des PEC a été évalué. Les résultats ont montré que les méthodes d'extraction et le pH de la solution extraitante ont influencé la composition des PEC et leur hydrophobicité. En outre, les extraitants des PEC, par exempe le formaldéhyde, l'éthanol, le dodécylsulfate de sodium (SDS) et Tween 20, ont non seulement introduit une teneur supplémentaire en carbone pendant la mesure du carbone organique total (COT), mais ils ont également interagit avec la résine DAX-8. En comparant la répartition du poids moléculaire apparent (aMW) des échantillons des PEC non traités et ajustés au pH détectés par chromatographie d'exclusion stérique (en anglais SEC), l’information plus complète d’aMW a été préservée à pH 5. Ainsi, le fractionnement hydrophobe par la résine DAX-8 à pH 5 et les méthodes physiques d'extraction PEC ont été préférés dans cette étude.Une analyse qualitative détaillée des caractéristiques hydrophobes des EPS a été étudiée par la technique de la matrice de fluorescence d’excitation-emission (EEM). Les résultats ont montré que les substances de type humique (HS-like) représentaient la majorité des composés organiques des PEC extraits de la boue granulaire anaérobie, et constituaient également le principal support moléculaire de l'hydrophobicité des extraits. Ces composés hydrophobes de type HS étaient essenciellement des molécules petites tailles de 8 kDa à <1 kDa. L’hydrophobité contributée par les protéines (PN) et les polysaccharides (PS) présentait un moindre rapport.Afin d’explorer les propriétés hydrophobes de PN et de PS, ainsi évaluer l'impact de l'addition de Ni(II) sur l'hydrophobicité des extraits des champignons, fongi Phanerochaete chrysosporium a été choisi. Les résultats ont montré que la teneur de PN et de PS dans les PEC extrait de ce type de fongi variait en fonction de la concentration de Ni(II). Avec une augmentation de la concentration de Ni de 0 mg/L à 25 mg/L, la teneur en PN a diminué alors que celle de PS a été augmentée. L'hydrophobicité des PEC du fongi, déterminée par le traitement de la résine DAX-8, a diminué lors que la concentration de Ni augmentait. Par ailleurs, l'intensité du pic de SEC correspondant aux molécules PN-like (Ex/Em = 225/345 nm) de 1,9 × 103 à 10 kDa a été augmentée par l'addition Ni; en même temps, la distribution d’aMW des composés organiques totaux (UV/210) dans les PEC restait presque stable. Ces résultats ont indiqué que les composés de type PN-like peuvent avoir déterminé l'hydrophobicité des PEC fongique dans des conditions de stress.Dans l’extrait plus hétérogène des PEC de boues granulaires anaérobies, des composés HS-like représentaient le composant organique majeur, ainsi le principal support moléculaire de l'hydrophobicité des PEC. En étudiant les caractéristiques hydrophobes des PEC extrait du champignon Phanerochaete chrysosporium, le PN et le PS des PEC jouaient un rôle actif dans la protection du champignon sous le Ni. La concentration élevée de Ni a diminué l'hydrophobicité des PEC fongique, mais elle a augmenté l'hydrophobicité de la surface cellulaire du champignon. Il semble que la présence de Ni favorise l'apparition d'un champignon plus hydrophobe / The hydrophobic properties of extracellular polymeric substances (EPS) exert a profound influence on the cell surface properties. However, many factors such as EPS extractions methods, substrate type influence EPS characteristics, and limited information regarding to the hydrophobic features of EPS can be found. The main aim of this study is to develop a proper method to study EPS hydrophobicity, and then investigate the hydrophobic features of EPS.The hydrophobic fractionation by Supelite™ DAX-8 resin was first applied on the EPS extracted from anaerobic granular sludge, two elution pH conditions i.e. pH 2 and 5 were tested. The impact of seven EPS extraction methods on the hydrophobic features of EPS was assessed. The results showed that the extraction methods and bulk solution pH dramatically influenced the EPS composition and their hydrophobicity. Besides, the EPS extracting reagents namely formaldehyde, ethanol, sodium dodecyl sulfate (SDS) and Tween 20 not only introduced extra carbon content during total organic carbon (TOC) measurement, but also interacted with the DAX-8 resin. By comparing the apparent molecular weight (aMW) distribution of the untreated and pH-adjusted EPS samples detected by size exclusion chromatography, more complete EPS aMW information was preserved at pH 5. Thus, the hydrophobic fractionation by DAX-8 resin at pH 5 and physical EPS extraction methods were preferred in this study.After identifying the proper conditions for DAX-8 resin fractionation, detailed qualitative analysis of the EPS hydrophobic features was further investigated. The results showed that the humic-like substances (HS-like) were the major organic constituent of the EPS extracted from the anaerobic granular sludge, and they were also the main molecular support of the EPS hydrophobicity. Those hydrophobic HS-like compounds were mainly small molecules ranging from 8 kDa to <1 kDa. Proteins (PN) and polysaccharides (PS) contributed to the EPS hydrophobicity to a lesser extent.The role of PN and PS in the EPS hydrophobicity was difficult to be shown. It is known that the major organic constituents of the EPS extracted from bacteria, algae and fungi are PN and PS. Therefore, to explore the hydrophobic features of PN and PS, as well as to investigate the impact of Ni(II) addition, on the EPS hydrophobicity, the fungus Phanerochaete chrysosporium was chosen. The results showed that the contents of PN and PS in the extracted fungal EPS varied with the Ni(II) concentration. With an increase in the Ni concentration from 0 mg/L to 25 mg/L, the PN content was decreased whereas the PS content was increased. The fungal EPS hydrophobicity, determined by the DAX-8 resin treatment, was decreased as the Ni concentration increased.Besides, the peak intensity on the size exclusion chromatography (SEC) corresponding to the PN-like molecules (Ex/Em = 225/345 nm) ranging from 1.9×103 to 10 kDa were intensified by the Ni addition, while the aMW distribution of the total organics (UV/210) in the EPS remained almost stable. These results indicated that those PN-like compounds may determine the hydrophobicity of fungal EPS under stress conditions.For the more heterogeneous EPS extracted from anaerobic granular sludge, HS-like compounds were identified as the major organic component, as well as the main molecular support of the EPS hydrophobicity. By studying the hydrophobic features of the EPS extracted from the fungus Phanerochaete chrysosporium, it showed that the PN and PS in the EPS played an active role in protecting the fungus under Ni stress. The increased Ni concentration decreased the hydrophobicity of fungal EPS, but it increased the cell surface hydrophobicity of the fungus. It seems that the presence of Ni promoted the fungus becoming more hydrophobic

Corantes reativos têxteis podem ser degradados por basidiomicetos, por meio de enzimas oxidativas e hidrolíticas, e compostos de baixa massa molar. Foi avaliada a descoloração de CI Reactive Blue 222 por Peniophora cinerea, Pleurotus ostreatus e Trametes villosa, selecionando condições ótimas para o processo e diferentes vias metabólicas foram observadas. A degradação foi confirmada por cromatografia de camada delgada. Foi sugerido que lacases de P. ostreatus oxidam o grupo cromóforo azo, ligado ao fenol, nas primeiras 24 horas, conjuntamente hidroxilização não enzimática. Lacases de P. cinerea oxidam Mn+2 e quinona, possibilitando a via de Fenton e hidroxilizando assim a molécula do corante, paulatinamente, a partir das ligações mais vulneráveis. T. villosa faz uso prioritariamente da via de Fenton, hidroxilizando gradativamente a molécula do corante. Dessa forma, embora a maioria de estudos associem a produção enzimática à descoloração, a participação dos compostos de baixa massa molar não pode ser negligenciada. / Reactive textile dyes can be degraded by basidiomycetes, by means of hydrolytic and oxidative enzymes, and low molecular weight compounds. Was evaluated the CI Reactive Blue 222 decolorization by Peniophora cinerea, Pleurotus ostreatus, and Trametes villosa, selecting optimal conditions for the process and different metabolic pathways were observed. The degradation was confirmed by thin layer chromatography. It was suggested that P. ostreatus laccases oxidize azo chromophore group attached to the phenol, within 24 hours, together nonenzymatic hydroxylizating. P. cinerea laccases oxidize Mn+2 and quinone, enabling via Fenton and so hidroxylizing the dye molecule, gradually, from the most vulnerable links. T. villosa uses primarily via Fenton, gradually hidroxylizing the dye molecule. Thus, although most studies have linked enzyme production with the decolorization, the share of low molecular weight compounds can not be neglected.

Physical aging of commercially produced polypropylene homopolymers Mosten was studied. The work is focused on monitoring the changes in physical properties of polymer in the time period 90 days from the date of preparation of the test specimen. The aging of polymers was realized at storage temperatures of 23 and 80 °C. The changes in crystallinity, yield strength, modulus, elongation and impact strength using DSC method, tensile tests and impact tests according to Charpy were monitored. Molecular weight distribution was determined by means of GPC at each polymer. Relationships between the changes in physical properties and polymer molecular weight were sought.

Herein, we present a quasi-living Negishi-type catalyst-transfer polycondensation of a zinc–organic DTSbased monomer which provides an access to narrowly distributed poly(4,4-bis(2-ethylhexyl)dithieno[3,2-b:20,30-d]silole (PDTS) with controlled molecular weight. The synthesis of well-defined all-conjugated diblock copolymers containing a PDTS block was accomplished by a combination of Kumada and Negishi catalyst-transfer polycondensations (KCTP and NCTP, respectively). Particularly, it was shown that living P3HT chains obtained by KCTP of magnesium–organic thiophene-based monomer efficiently initiate NCTP of zinc–organic DTS-based monomer. The purity of the DTS-based monomer was found to be a crucial factor for achieving a clean chain-growth polymerization process. A combination of physico-chemical methods was used to prove the success of the block copolymerization.

CHARLES UNIVERSITY IN PRAGUE FACULTY OF SCIENCE DEPARTMENT OF BIOCHEMISTRY PRAGUE 2010 Supervisor: Doc. MUDr. Jiří Gallo, PhD. Supervisor-consultant: Prof. RNDr. Gustav Entlicher, CSc. Eva Zolotarevová Summary of PhD Thesis POLYETHYLENE WEAR PARTICLES AROUND TOTAL JOINT REPLACEMENTS - THEIR PROPERTIES, DISTRIBUTION AND POSSIBLE MECHANISM OF THEIR ADVERSE BIOLOGICAL EFFECTS PRAGUE 2010 The research project was supported by the Grant project of National research program II. (no. 2B06096) of the Czech Ministry of Education, Youth and Sports named "Observation and minimization of UHMWPE wear in joint replacements". Front-page picture: Polyethylene wear particle isolated from granuloma tissue surrounding total hip replacement; Electron microscope microphotograph; particle size app. 90 x 80 µm; retouched Eva Zolotarevová Introduction Introduction For many people all over the world a replacement with an implant of irreversibly damaged joint is often the only way how to come back to normal life without pain and mobility limitation. Nowadays, greater and greater requirements are asked of quality of all replacement components, especially in terms of quality of used materials. Biomechanical properties of hip and knee replacements have been already solved. The most often used materials for manufacturing joint...

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While a decrease of wild felid population has led to disruption of conservation programme, recent studies have shown the importance of immune regulation for determining health outcomes and co-infection. Immunoglobulin G is important for detecting and evaluating responses to infectious diseases and vaccination. But, there is limited information on felid immunoglobulins and their role for functional immunity. This study aimed at isolating and characterizing lion’s immunoglobulin G. Lions’ sera (n = 68) were processed using the MagReSyn® magnetic beads and the final protein concentration was determined using the Xpose™ Trinean Spectrophotometer. The cross-reactivity of goat anti-cat immunoglobulin with sera of lions and other species was analysed using ELISA. High cross-reactivity was observed in lions ranging from 87.7 to 100%, and low reactivity with rhino (22.4%) followed by chicken (0.01%). The protein concentration from purified sera yielded 39.09 mg/ml. Molecular weight of lion IgG 150-160 kDa was detected with both chains at 54-56 kDa and 24-26 kDa on SDS PAGE. These results indicate a potential aid in developing serological tools to monitor exposure to micro-organisms of lions. / Agriculture and  Animal Health / M. Sc. (Agriculture)